acid-phosphatase and Leukemia--Lymphocytic--Chronic--B-Cell

Bone destruction, caused by aberrant production and activation of osteoclasts, is a prominent feature of multiple myeloma. We demonstrate that myeloma stimulates osteoclastogenesis by triggering a coordinated increase in the tumor necrosis factor-related activation-induced cytokine (TRANCE) and decrease in its decoy receptor, osteoprotegerin (OPG). Immunohistochemistry and in situ hybridization studies of bone marrow specimens indicate that in vivo, deregulation of the TRANCE-OPG cytokine axis occurs in myeloma, but not in the limited plasma cell disorder monoclonal gammopathy of unknown significance or in nonmyeloma hematologic malignancies. In coculture, myeloma cell lines stimulate expression of TRANCE and inhibit expression of OPG by stromal cells. Osteoclastogenesis, the functional consequence of increased TRANCE expression, is counteracted by addition of a recombinant TRANCE inhibitor, RANK-Fc, to marrow/myeloma cocultures. Myeloma-stroma interaction also has been postulated to support progression of the malignant clone. In the SCID-hu murine model of human myeloma, administration of RANK-Fc both prevents myeloma-induced bone destruction and interferes with myeloma progression. Our data identify TRANCE and OPG as key cytokines whose deregulation promotes bone destruction and supports myeloma growth.

Topics: Acid Phosphatase; Animals; Carrier Proteins; Disease Progression; Glycoproteins; Hodgkin Disease; Humans; Isoenzymes; Leukemia, Lymphocytic, Chronic, B-Cell; Membrane Glycoproteins; Mice; Mice, SCID; Osteoprotegerin; Paraproteinemias; RANK Ligand; Receptor Activator of Nuclear Factor-kappa B; Receptors, Cytoplasmic and Nuclear; Receptors, Tumor Necrosis Factor; Reference Values; Reverse Transcriptase Polymerase Chain Reaction; Tartrate-Resistant Acid Phosphatase; Time Factors

We have previously reported that bryostation 1 (Bryo 1) induces differentiation of chronic lymphocytic leukemia (CLL) in vitro to a hairy cell (HC) stage. This study tests the hypothesis that Bryo 1-differentiated CLL cells are more susceptible to 2-chlorodeoxyadenosine (2-CdA) than parent CLL cells. A recently established EBV-negative CLL line (WSU-CLL) from a patient resistant to chemotherapy including fludarabine was used to test this hypothesis. Both Bryo 1 (10-1000 nM) and 2-CdA (5.6-22.4 microM) exhibited a dose-dependent growth inhibitory effect on the WSU-CLL cell line. In vitro, the sequential exposure to Bryo 1 (100 nM for 72 h) followed by 2-CdA (11.2 microM) resulted in significantly higher rates of growth inhibition than either agent alone. Changes in immunophenotype, enzymes, lipids, proteins, and the DNA of WSU-CLL cells were studied before and after Bryo 1 treatment. Bryo 1 induced a positive tartrate-resistant acid phosphatase reaction and two important markers, CD11c and CD25, after 72 h of culture, confirming the differentiation of CLL to HC. The Fourier transformation infrared spectroscopic analysis showed that the amount of membrane lipids significantly increased in Bryo 1-treated cells compared to controls after 24 h, whereas the protein content, as well as the DNA content, decreased. This finding supports the change of CLL to HC. To evaluate the in vivo efficacy of Bryo 1 and 2-CdA, we used a xenograft model of CLL in WSU-CLL-bearing mice with severe combined immune deficiency. s.c. tumors were developed by injection of 10(7) WSU-CLL cells, and fragments were then transplanted into a new batch of severe combined immunodeficient mice. Bryo 1 and 2-CdA at the maximum tolerated doses (75 micrograms/kg i.p. and 30 mg/kg s.c., respectively) were administered to the mice at different combinations and schedules. The survival in days, the tumor growth inhibition ratio, the tumor growth delay, and the log10 kill of the mice treated with Bryo 1 followed by 2-CdA were significantly better than the control and other groups. We conclude that the sequential treatment with Bryo 1 followed by 2-CdA resulted in higher antitumor activity and improved animal survival.

Topics: Acid Phosphatase; Aged; Animals; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Bryostatins; Cell Division; Cladribine; DNA, Neoplasm; Drug Administration Schedule; Drug Screening Assays, Antitumor; Humans; Lactones; Leukemia, Hairy Cell; Leukemia, Lymphocytic, Chronic, B-Cell; Lipid Metabolism; Macrolides; Male; Mice; Mice, SCID; Neoplasm Proteins; Neoplasm Transplantation; Spectroscopy, Fourier Transform Infrared; Transplantation, Heterologous; Tumor Cells, Cultured

The demonstration of tartrate-resistant acid phosphatase (TRAP) activity has long been a cornerstone in the diagnosis of hairy cell leukemia (HCL). Recently a monoclonal antibody to this enzyme has been developed that can be used in an immunoperoxidase method on paraffin-embedded tissues. By using a peroxidase-labeled streptavidin biotin method, paraffin sections of B5 and formalin-fixed tissue from 86 cases of HCL (41 bone marrow, 36 spleen, 9 liver) were stained with the antibody to TRAP and compared against staining for CD20 (L26) and DBA.44 (DAKO, Carpinteria, Calif). In addition, 193 specimens (127 bone marrow, 42 lymph node, 19 spleen, 5 other) from a variety of neoplastic and nonneoplastic hematologic conditions were stained using the monoclonal antibody to TRAP. For comparison, these cases were also stained with DBA.44. In the cases of HCL, 80 of 86 specimens were immunoreactive for TRAP. While the antibody to TRAP generally stained less than 50% of the hairy cells, CD20 and DBA.44 stained 90% and 50% to 60% of hairy cells, respectively. Two of three cases of marginal zone lymphoma showed weak immunoreactivity to the TRAP antibody. Two specimens from a patient with Gaucher's disease and 8 of 13 cases of mastocytosis also showed positivity to the TRAP antibody in the macrophages and mast cells, respectively. In contrast, staining for DBA.44 was positive in 3 of 9 cases of B-cell large cell lymphoma, 1 of 4 cases of mantle cell lymphoma, and in the paraimmunoblasts of 1 of 7 cases of small lymphocytic lymphoma. Only HCL was TRAP and DBA.44 positive. This antibody to TRAP is a useful addition to the diagnosis of HCL but should be used in conjunction with CD20 and DBA.44. The use of this antibody to determine minimal residual disease after chemotherapy was not addressed.

Topics: Acid Phosphatase; Antibodies, Monoclonal; Biomarkers, Tumor; Bone Marrow; Bone Marrow Neoplasms; Diagnosis, Differential; Gaucher Disease; Humans; Immunohistochemistry; Isoenzymes; Leukemia, Hairy Cell; Leukemia, Lymphocytic, Chronic, B-Cell; Liver; Liver Neoplasms; Lymph Nodes; Lymphoma, B-Cell; Lymphoproliferative Disorders; Macrophages; Mast Cells; Paraffin Embedding; Pathology, Clinical; Spleen; Splenic Neoplasms; Tartrate-Resistant Acid Phosphatase

B-Chronic lymphocytic leukemia (B-CLL) and hairy cell leukemia (HCL) are both differentiated B-cell lymphoproliferative disorders. Prior studies have suggested that phorbol esters and the macrocyclic lactone Bryostatin-1, which are both protein kinase-C activators, can induce the differentiation of B-CLL cells into HCL cells in vitro, as evidenced by morphology, phenotype and TRAP activity. The differentiating effect of all-trans retinoic acid on B-CLL cells has been less extensively studied. We studied the effects of incubating adherence purified B-CLL cells with phorbol myristic acetate (PMA), all-trans retinoic acid (ATRA), and Bryostatin-1. None of these agents induced a true HCL phenotype (CD5-, CD11c/CD25 coexpression) under the conditions studied.

Topics: Acid Phosphatase; Antigens, CD; Antigens, Neoplasm; B-Lymphocytes; Biomarkers, Tumor; Bryostatins; Cell Differentiation; Enzyme Activation; Humans; Immunophenotyping; Isoenzymes; Lactones; Leukemia, Hairy Cell; Leukemia, Lymphocytic, Chronic, B-Cell; Macrolides; Neoplastic Stem Cells; Protein Kinase C; Tartrate-Resistant Acid Phosphatase; Tetradecanoylphorbol Acetate; Tretinoin; Tumor Cells, Cultured

We investigated the effects of stromal cells, obtained from lymph nodes with various lymphoid disorders and characterized immunocytochemically as fibroblasts, on the maturation of a cell line "HN" established from an atypical PLL (prolymphocytic leukemia) having phenotypic characteristics of both CLL (chronic lymphocytic leukemia) and HCL (hairy cell leukemia). Coculture with stromal cells, irrespective of the kind of lymphoid disease affecting the lymph node, induced a plasmacytoid cytology in HN cells, an increase of cIg gamma, and decreases of sIg gamma, CD5, CD21, HC2, and B-ly7. In contrast, coculture with the stromal cells produced no marked effects on hairy cells from two HCL patients. Similarly, coculture with stromal cell supernatant or with various cytokines shown to be produced by bone marrow stromal cells produced no effects on HN cells. We propose that lymph node stromal cells play an important role in the differentiation of B cells through direct contact and that HN cells will be useful for investigating the differentiation pathway of chronic B cell leukemia cells.

Topics: Acid Phosphatase; Antigens, CD; B-Lymphocytes; Blood Coagulation Factors; Cell Differentiation; Cells, Cultured; Cytokines; Humans; Leukemia, Hairy Cell; Leukemia, Lymphocytic, Chronic, B-Cell; Leukemia, Prolymphocytic; Lymph Nodes; Lymphatic Diseases; Phenotype; Receptors, Antigen, B-Cell; Stromal Cells; Tumor Cells, Cultured

Peripheral blood mononuclear cells from 11 patients with untreated B-chronic lymphocytic leukemia (CLL) were exposed to sodium phenylacetate (NaPA) in culture to assess its ability to induce differentiation. We found no evidence of cellular differentiation or induction of tartrate resistant acid phosphatase activity, as seen when B-CLL cells were treated with phorbol ester. We observed a striking decrease in the viability of the B-CLL cells in a time and dose dependent fashion when exposed to NaPA. After six days of culture, control cells from the 11 patients studied had a median viability of 90%, whereas cells exposed to NaPA at 5 and 10 mM concentrations had median viabilities of 39 and 16%, respectively. The cells treated with NaPA developed prominent cytoplasmic vacuoles. NaPA binds and depletes glutamine which is an important amino acid for lymphocyte metabolism. Although the mechanism of the cytocidal effects demonstrated in this study are unknown, they may relate at least partially to glutamine deprivation.

Topics: Acid Phosphatase; B-Lymphocytes; Cell Differentiation; Cell Survival; Drug Screening Assays, Antitumor; Enzyme Induction; Gene Expression Regulation, Leukemic; Humans; Leukemia, Lymphocytic, Chronic, B-Cell; Neoplasm Proteins; Neoplastic Stem Cells; Palatine Tonsil; Phenylacetates; Tumor Cells, Cultured

The phorbol esters induce differentiation of chronic lymphocytic leukemia (CLL) cells. Clinical use of this observation has been hampered by the fact that phorbol esters are also tumor promoters. In this study we demonstrate that another protein kinase C activator, without tumor promoting activity, has similar effects on CLL cells. Fresh leukemic cells from the peripheral blood of 13 patients with CLL were isolated and cultured in the absence (control) or presence of Bryostatin 1 or 12-0-tetradecanoylphorbol 13-acetate (TPA). Aliquots of cells were then analyzed after 24, 72 and 120 hours for morphological changes, acid phosphatase (ACP) and the co-expression of two hairy cell-associated surface antigens, CD22 and CD11c, by flow cytometry. Bryostatin 1 induced changes in shape and morphology similar to TPA, with adherence and increase in cell size, abundant cytoplasm and irregular cytoplasmic membrane. Both agents induced a statistically significant increase in the expression of CD22 and CD11c compared with control (p < 0.0008). There was no significant difference between the two agents in the degree of expression of these two markers. Both agents also induced ACP that was tartrate resistant (TRAP). These changes indicate that Bryostatin is as effective as TPA in inducing further differentiation of CLL cells to a hairy cell stage.

Topics: Acid Phosphatase; Aged; Aged, 80 and over; Antigens, CD; Antigens, Differentiation, B-Lymphocyte; Bryostatins; CD11 Antigens; Cell Adhesion Molecules; Cell Differentiation; Enzyme Activation; Female; Humans; Immunophenotyping; Lactones; Lectins; Leukemia, Hairy Cell; Leukemia, Lymphocytic, Chronic, B-Cell; Macrolides; Male; Middle Aged; Protein Kinase C; Sialic Acid Binding Ig-like Lectin 2; Tetradecanoylphorbol Acetate; Tumor Cells, Cultured

Leukemic cells from eight patients with chronic lymphocytic leukemia were isolated and cultured in the continuous presence of 12-O-tetradecanoylphorbol 13-acetate (TPA) at a concentration of 1.6 x 10(-9) M for 4-10 days. Aliquots of cells were then analyzed at intervals of 24-72 hr for changes in morphology, acid phosphatase staining (AP), and expression of two hairy cell-associated surface antigens, HCL1 (CD22, Leu 14) and HCL3 (CD11c, Leu M5). All cases studied showed typical B-CLL phenotype, and only a small proportion of cells expressed CD22 and CD11c (mean 7% and 4.9%, respectively). TPA treatment induced the coexpression of CD22 (mean 49%) and CD11c (mean 48%) and tartrate-resistant acid phosphatase in seven of eight cases. Morphologically, cells in TPA cultures expressed hairy cell features that were evident in light and electron microscopic studies. Collectively these changes indicate that TPA can induce hairy cell features on CLL cells in vitro, suggesting the later maturational stage of HCL compared with CLL.

Topics: Acid Phosphatase; Aged; Antigens, CD; Antigens, Differentiation, B-Lymphocyte; CD11 Antigens; Cell Adhesion Molecules; Dose-Response Relationship, Drug; Electrophoresis, Polyacrylamide Gel; Female; Humans; Lectins; Leukemia, Hairy Cell; Leukemia, Lymphocytic, Chronic, B-Cell; Lymphocytes; Male; Microscopy, Electron; Middle Aged; Phenotype; Phorbol Esters; Precipitin Tests; Sialic Acid Binding Ig-like Lectin 2; Tetradecanoylphorbol Acetate; Time Factors

Topics: Acid Phosphatase; Antigens, CD; CD11 Antigens; CD5 Antigens; Humans; Leukemia, Lymphocytic, Chronic, B-Cell; Leukemia, Prolymphocytic; Tartrates

Chronic lymphocytic leukemia (CLL) and hairy cell leukemia (HCL) are two common chronic lymphoproliferative disorders, each having characteristic clinical, morphologic, and immunologic features. Phenotypically, CD5 reactivity in CLL and CD11c (Leu-M5) reactivity in HCL have characterized these two leukemias among B-cell disorders. In this study, we report 14 cases of a novel chronic lymphoproliferative disorder characterized by lymphocytosis and CD11c expression, but morphologically similar to CLL. The patients' ages ranged from 46 to 81 years (median 62). Eleven had palpable splenomegaly, five with markedly enlarged spleens; only one patient had generalized lymphadenopathy. The white blood cell count ranged from 5.2 to 131.0 x 10(9)/L (median 20.8). The morphologic diagnosis in all cases was CLL, with the cells usually having abundant cytoplasm. No morphologic features, of hairy cells were evident; tartrate-resistant acid phosphatase cytochemistry was negative in all cases. Bone marrow biopsies were available in 8 of 14. Four showed focal nodular infiltrates and two had diffuse infiltrates similar to CLL; two showed only minimal interstitial involvement. All cases expressed multiple B-cell markers, and 12 of 14 had monoclonal surface immunoglobulin. The leukemic cells of all cases strongly expressed CD11c, while CD5 was expressed in 7 of 14; only 1 of the 14 cases expressed the lymph node homing receptor, Leu-8. This unique group of leukemias appears to represent the malignant transformation of lymphocytes arising from a stage of lymphocyte differentiation between that found in typical cases of CLL and that of HCL. CD11c is known to have an important function in cellular adhesion and may be important in determining the pattern of lymphocyte tissue distribution found in this group of patients.

Topics: Acid Phosphatase; Aged; Aged, 80 and over; Antigens, CD; Antigens, Differentiation; Antineoplastic Agents; B-Lymphocytes; Bone Marrow; CD11 Antigens; CD5 Antigens; Female; Histocytochemistry; Humans; Immunophenotyping; Leukemia, Hairy Cell; Leukemia, Lymphocytic, Chronic, B-Cell; Leukocytosis; Male; Middle Aged; Splenomegaly; Thrombocytopenia

Plasmacytic morphologic characteristics are usually associated with cells of B-lymphocyte origin. Recently, plasmacytoid T-cells have been described in reactive lymph nodes and a rare form of lymphoma characteristically associated with myeloproliferative disorders. This report documents a case of plasmacytoid T-cell malignancy that initially presented as an acute leukemia in an elderly man with a longstanding myelodysplastic syndrome. The tumor replaced bone marrow and involved lymph nodes. Despite aggressive therapy, he died quickly of his leukemia/lymphoma. This case illustrates the need for complete cellular analysis in the diagnosis of morphologically plasmacytic malignancies and raises additional questions about the relationship of this peculiar type of T-cell to the hematopoietic marrow.

Topics: Acid Phosphatase; Aged; Antibodies, Monoclonal; Antigens, CD; Cell Cycle; Diagnosis, Differential; DNA, Neoplasm; Fluorescent Antibody Technique; Gene Rearrangement, beta-Chain T-Cell Antigen Receptor; Histocytochemistry; Humans; Karyotyping; Leukemia, Lymphocytic, Chronic, B-Cell; Leukemia, Plasma Cell; Male; Microscopy, Electron; Myelodysplastic Syndromes; T-Lymphocytes

Cells from 3 patients with B-chronic lymphocytic leukaemia (B-CLL) and 1 with B-prolymphocytic leukaemia (B-PLL) were treated in vitro with the phorbol ester 12-0-tetradecanoylphorbol (TPA), the calcium ionophore A23187, or a combination of TPA and A23187. TPA induced the cells to adhere to the culture flask or to clump in dense clusters; single cells became enlarged, often with cytoplasmic elongations. Cells treated with TPA plus A23187 acquired a plasmacytoid morphology and formed regular aggregates in culture. Only TPA alone induced the expression of tartrate-resistant acid phosphatase (TRAP) as documented by isoelectric focusing on horizontal thin-layer gels. The TRAP isoenzyme was first detected after 24 h of TPA treatment; its intensity increased during further TPA exposure, being maximally expressed at 72/96 h. The results suggest that, while TPA triggers B-CLL cells to convert to hairy cell leukaemia (HCL)-type cells, the double stimulus which more closely imitates physiological activation initiates a 'normal' differentiation programme which leads to plasma cells.

Topics: Acid Phosphatase; Aged; B-Lymphocytes; Calcimycin; Drug Synergism; Female; Humans; Isoenzymes; Leukemia, Lymphocytic, Chronic, B-Cell; Leukemia, Prolymphocytic; Male; Middle Aged; Tartrates; Tetradecanoylphorbol Acetate